TWI608006B - Novel crystalline form - Google Patents

Description

Novel crystalline form

The present invention relates to epithelial sodium channel (ENaC) blocker 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1 , a novel crystalline form of succinic acid salt of 3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester, A pharmaceutical composition comprising the crystalline form, the crystalline form, and the use of the pharmaceutical compositions, and a method of preparing the crystalline form.

International Patent Application Publication No. WO 2012/035158 (PCT/EP2011/066151) discloses the preparation of the ENaC blocker 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine- 2-carbonylimino]-1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester And its succinate method.

Diseases mediated by blocking epithelial sodium channels include diseases associated with fluid volume regulation across the epithelial membrane. For example, the volume of fluid on the surface of the respiratory tract is a key regulator of mucociliary clearance and maintenance of lung health. Blocking the epithelial sodium channel will promote fluid accumulation on the mucosal side of the airway epithelium, thereby promoting mucus clearance and preventing mucus and saliva from accumulating in respiratory tissues, including the lung respiratory tract. Such diseases include respiratory diseases such as cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory infections (acute and chronic; viral and bacterial) And lung cancer. In addition to respiratory diseases, diseases mediated by blocking epithelial sodium channels also include abnormal fluid regulation through the epithelium and may be involved in the abnormality of protective surface fluids on their surface. Physiological diseases such as xerostomia (dry mouth) or dry keratoconjunctivitis (keratoconjunctivitis sire) (dry eye syndrome). In addition, blocking the epithelial sodium channel in the kidney can be used to promote diuresis and thereby induce a hypotensive effect.

Figure 1 shows the pattern for the A (Example 3) recording the x- ray powder diffraction pattern using CuKα radiation and a line (λ = 1.5418Å) recorded on a Bruker ^TM D8 diffractometer.

Figure 2 shows the differential scanning calorimetry (DSC) trace of Form A (Example 3) and recorded on a Perkin Elmer Diamond DSC instrument (Perkin Elmer, model BO14-3018) with an aluminum pan; heating rate 20 K/min, Temperature range: 30 ° C to 250 ° C.

Figure 3 shows the infrared spectrum of Form A (Example 3) and was recorded on a Bruker Alpha instrument in an attenuated total reflectance (ATR) mode. The measurement parameters are as follows: range 400 cm ^-1 to 4000 cm ^-1 , resolution 2 cm ^-1 , number of scans 64, speed 7.5 kHz, apodization method: Blackman-Harris 3-term.

Figure 4 shows the pattern for the A (Examples 1,2) x- ray powder diffraction pattern of the recording system and using CuKα radiation (λ = 1.5418Å) recorded on a Bruker ^TM D8 diffractometer.

Figure 5 shows the differential scanning calorimetry (DSC) trace of Form A (Examples 1, 2) and recorded on a Perkin Elmer Diamond DSC instrument (Perkin Elmer, model BO14-3018) with an aluminum pan; heating rate 20 K/ Min, temperature range: 30 ° C to 250 ° C.

Figure 6 shows the pattern for the B (Examples 4,5,6,7) x- ray powder diffraction pattern of the recording system and using CuKα radiation (λ = 1.5418Å) recorded on a Bruker ^TM D8 diffractometer.

Figure 7 shows the pattern for the C (Example 10) x- ray powder diffraction pattern of the recording system and using CuKα radiation (λ = 1.5418Å) recorded on a Bruker ^TM D8 diffractometer.

Figure 8 shows the differential scanning calorimetry (DSC) trace of Form C (Example 10) and recorded on a Perkin Elmer Diamond DSC instrument (Perkin Elmer, model BO14-3018) with an aluminum pan; heating rate 20 K/min, Temperature range: 30 ° C to 250 ° C.

Figure 9 shows the infrared spectrum of Form C (Example 10) and was recorded on a Bruker Alpha instrument in an attenuated total reflectance (ATR) mode. The measurement parameters are as follows: range 400 cm ^-1 to 4000 cm ^-1 , resolution 2 cm ^-1 , number of scans 64, speed 7.5 kHz, apodization method: Blackman-Harris 3-term.

Figure 10 shows the pattern for the C (Examples 8,9) x- ray powder diffraction pattern of the recording system and using CuKα radiation (λ = 1.5418Å) recorded on a Bruker ^TM D8 diffractometer.

Figure 11 shows the DSC trace of Form C (Examples 8, 9) and recorded on a Perkin Elmer Diamond DSC instrument (Perkin Elmer, Model BO14-3018) with an aluminum pan; heating rate 20 K/min, temperature range: 30 ° C to 250 ° C.

Revealing ENaC Blocker 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza - Three polymorphic forms of succinates of spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester (forms A, B and C) (3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[4.5 ] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl ester: succinic acid = 1:1). The method of preparing the types A and B is described in International Patent Application Publication No. WO 2012/035158 (PCT/EP2011/066151).

Thus, in a first aspect, the invention provides 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino] of the formula Crystalline form of succinate salt of -1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester ,

It contains version C.

In one embodiment of the first aspect, the crystalline form consists essentially of Form C.

In another embodiment of the first aspect, the crystalline form comprises Form C, wherein the Form C is in a substantially pure form.

In another embodiment of the first aspect, the crystalline pattern is characterized in that the x-ray powder diffraction pattern comprises 4 or more 2-theta values selected from the group consisting of: 21 ° C to 26 ° C : 7.0°±0.2, 10.6°±0.2, 14.3°±0.2, 18.2°±0.2, 18.6°±0.2, 19.2°±0.2, 21.2°±0.2, 21.8°±0.2, 24.7°±0.2, 29.0°±0.2 And 31.5 ° ± 0.2.

In another embodiment of the first aspect, the crystalline pattern is characterized in that the x-ray powder diffraction pattern comprises 6 or more 2-theta values selected from the group consisting of: 21 ° C to 26 ° C : 7.0°±0.2, 10.6°±0.2, 14.3°±0.2, 18.2°±0.2, 18.6°±0.2, 19.2°±0.2, 21.2°±0.2, 21.8°±0.2, 24.7°±0.2, 29.0°±0.2 And 31.5 ° ± 0.2.

In another embodiment of the first aspect, the x-ray powder diffraction spectrum of the crystalline form is substantially the same as the x-ray powder diffraction spectrum shown in FIG.

In another embodiment of the first aspect, the differential scanning calorimetry thermogram of the crystalline version is substantially identical to the differential scanning calorimetry thermogram shown in FIG.

The term "substantially pure" as used herein when referring to a particular polymorphic form means that the polymorphic form comprises less than 10% by weight, preferably less than 5% by weight, more preferably less than 3% by weight, most preferably less than 1% by weight of the compound. Any other physical form (polymorph).

As used herein, "polymorph" refers to a crystalline form in which the molecules, atoms, and/or ions forming the crystal have the same chemical composition but differ in spatial arrangement.

Throughout this specification and the claims below, the words "comprise" or variations (such as "comprises" or "comprising") shall be understood to mean implied, unless the context requires otherwise. The integers or steps or integer groups or groups of steps are included, but do not exclude any other integer or step or group of integers or groups of steps.

The term "crystalline form of the invention" as used herein refers to a crystalline form as defined in any of the first or first aspects.

In a second aspect, a pharmaceutical composition comprising a crystalline form of any of the first or first aspects and a pharmaceutically acceptable carrier or diluent is provided.

In an embodiment of the second aspect, wherein Form C is in a substantially pure form.

In another embodiment of the second aspect, the pharmaceutical composition is in an inhalable form.

In a third aspect, a pharmaceutical composition of an embodiment combining a second or second aspect of one or more other active ingredients is provided.

In a fourth aspect, the crystalline form of any of the first aspect or the first aspect and the pharmaceutical composition of any of the second or second aspects are provided for treating a respiratory disease For example, cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory infections (acute and chronic; viral and bacterial) and lung cancer.

In a fifth aspect, the use of the crystalline form of any of the first aspect or the first aspect, or the composition of any of the second or second aspects, in the manufacture of a medicament, The medicament is for treating respiratory diseases such as cystic fibrosis and primary fibrosis Dysfunction, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory infections (acute and chronic; viral and bacterial) and lung cancer.

In a sixth aspect, there is provided a treatment for respiratory diseases (eg, cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory infections (acute and chronic; virus) Method of sexual and bacterial) and lung cancer comprising administering a crystalline form or a second or second aspect of any of the first or first aspects of the effective amount of the patient in need thereof A pharmaceutical composition of any of the embodiments.

In a seventh aspect, an inhalation device comprising a crystalline version of any of the embodiments in which the first aspect or the first aspect is delivered by pulmonary administration is provided. In certain preferred embodiments, the inhalation device is a dry powder inhaler, such as a BREEZHALER® inhaler.

3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro has been shown [ 4.5] The novel polymorphic form C of decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate is thermodynamically more stable than Form A.

Form B is a group of amorphous solvates which exhibit a transient crystalline form in suspension during the crystallization of Form A. Form B is converted to Form A upon drying.

The agent of the present invention may be administered by any suitable route, for example, orally, for example, in the form of a lozenge or capsule; parenterally, for example, intravenously; topically applied to the skin; intranasally, for example, In the treatment of allergic rhinitis; or preferably, by inhalation, especially in the treatment of obstructive or inflammatory respiratory diseases. In particular, the agents of the invention can be delivered as an inhalable formulation for the treatment of COPD, cystic fibrosis and asthma.

The pharmaceutical compositions can be formulated for specific routes of administration, such as oral administration, parenteral administration, and rectal administration. In addition, the pharmaceutical compositions of the present invention may be in solid form (including but not limited to, capsules, troches, pills, granules, powders or suppositories), or in liquid form (including but not limited to solutions, suspensions or emulsions) preparation. The pharmaceutical compositions may be subjected to conventional pharmaceutical operations (e.g., sterilization) and/or may contain conventional inert diluents, lubricants or buffers, and adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers and buffers, and the like.

In general, the pharmaceutical compositions comprise the active ingredient together with the following lozenge or gelatin capsules: a) diluents such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine; Lubricants, such as cerium oxide, talc, stearic acid, magnesium or calcium salts thereof and/or polyethylene glycol; for tablets also contain c) binders such as magnesium aluminum silicate, starch paste, gelatin , sulfonate, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone; optionally including d) a disintegrant such as starch, agar, alginic acid or a sodium salt thereof or an effervescent mixture; And / or e) absorbents, colorants, flavors and sweeteners.

Tablets can be film coated or enteric coated according to methods known in the art.

Compositions suitable for oral administration include an effective amount of a tablet of the invention in the form of a tablet, a lozenge, an aqueous or oily suspension, a dispersible powder or granule, an emulsion, a hard or soft capsule, or a syrup or elixir. Type. Compositions intended for oral use can be prepared according to any of the methods known in the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives. The agent is intended to provide a pharmaceutically elegant and palatable preparation. Tablets may contain a mixture of the active ingredient in admixture with a pharmaceutically acceptable pharmaceutically acceptable excipient. For example, such excipients are inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating agents and disintegrating agents, for example, corn starch or alginic acid; binders, for example, Starch, gelatin or gum arabic; and a lubricant such as magnesium stearate, stearic acid or talc. Such tablets are uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. Formulations for oral use can be presented in the form of hard gelatin capsules in which the active ingredient is combined with an inert solid diluent (for example, calcium carbonate, calcium phosphate or Mix; or may be provided in the form of a soft gelatin capsule in which the active ingredient is mixed with a water or oil medium (for example, peanut oil, liquid paraffin or olive oil).

Certain injectable compositions are isotonic aqueous solutions or suspensions, and suppositories are preferably prepared from fatty emulsions or suspensions. The compositions may be sterilized and/or contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, dissolution promoters, salts for regulating osmotic pressure, and/or buffers. In addition, it may also contain other therapeutically valuable substances. The compositions are prepared according to conventional mixing, granulating or coating methods and contain from about 0.1% to about 75%, or from about 1% to about 50%, by weight of active ingredient.

Compositions suitable for transdermal administration comprise an effective amount of a crystalline form of the invention and a suitable carrier. Carriers suitable for transdermal delivery include absorbable pharmaceutically acceptable solvents to aid passage through the host skin. By way of example, the transdermal device is in the form of a bandage comprising a backing member; a reservoir containing the compound, optionally with a carrier; optionally a speed control barrier to deliver the compound to the host for a prolonged period of time at a controlled predetermined rate Skin; and components that secure the device to the skin.

Compositions suitable for topical application (e.g., to the skin and eyes) include aqueous solutions, suspensions, ointments, creams, gels, or sprayable formulations such as delivered by aerosol or the like. Such topical delivery systems are especially useful for epidermal administration to, for example, treat skin cancer, for prophylactic use, for example, sunscreens, lotions, sprays, and the like. Thus, it is particularly suitable for topical application, including cosmetic formulations well known in the art. These formulations may contain solubilizers, stabilizers, penetration enhancers, buffers, and preservatives.

As used herein, topical application can also be by inhalation or intranasal administration. It may be in the form of a dry powder, either alone or in the form of a mixture of a dry blend and a lactose, or a mixed component particle with, for example, a phospholipid, with or without the use of a suitable propellant, from a dry powder inhaler, Easily delivered from a pressurized container, pump, ejector, nebulizer or nebulizer in aerosol spray delivery.

When the inhalable form of the active ingredient is an aerosol composition, the inhalation device can be a valved aerosol vial suitable for delivering a metered amount (eg, 10 μl to 100 μl, such as 25 μl to 50 μl) of composition, known as metered dose inhalation Device. Those skilled in the art of inhalation therapy are well aware of procedures suitable for such aerosol vials and for incorporating aerosol compositions into such vials under pressure. For example, the aerosol composition can be administered from a canister, for example as described in EP-A-0642992. When the inhalable form of the active ingredient is a sprayable aqueous, organic or aqueous/organic dispersion, the inhalation device can be a known nebulizer, such as a conventional pneumatic nebulizer, such as an air jet nebulizer or an ultrasonic nebulizer. Containing, for example, 1 ml to 50 ml, usually 1 ml to 10 ml dispersion; or hand-held spray, often referred to as a light mist or light spray inhaler, for example, an electronic control device such as AERx (Aradigm, US) or Aerodose (Aerogen) Or a mechanical device (such as a RESPIMAT (Boehringer Ingelheim) nebulizer that results in a reduced atomization volume (eg, 10 [mu]l to 100 [mu]l) compared to conventional nebulizers). When the inhalable form of the active ingredient is in the form of fine particles, the inhalation device can be, for example, adapted to deliver a dry powder from a capsule or blister containing a dry powder comprising unit doses (A) and/or (B). A dry powder inhalation device, or a multi-dose dry powder inhalation (MDPI) device suitable for delivering, for example, from 3 mg to 25 mg of dry powder comprising unit doses (A) and/or (B). The dry powder composition preferably contains a diluent or carrier such as lactose and a compound which helps prevent degradation of product properties due to moisture, such as magnesium stearate. Suitable dry powder inhaler device comprising the apparatus disclosed in the following: US 3991761 (including means AEROLIZER ^TM), WO 05/113042 (including means BREEZHALER ^TM), WO 97/20589 (including means CERTIHALER ^TM), WO 97/30743 ( comprising means TWISTHALER ^TM), WO 05/37353 (including means GYROHALER ^TM), US6536427 (including DISKUS ^TM device), WO 97/25086 (including means DISKHALER ^TM), WO 95/14089 (including means GEMINI ^TM), WO 03 / 77979 (including PROHALER ^(TM) devices), and devices disclosed in WO 08/51621, WO 09/117112, and US 2005/0183724.

The invention also includes (A) a crystalline form of the invention in an inhalable form; (B) comprising the present invention Inhalable form of a pharmaceutically acceptable form of the invention in inhalable form with a pharmaceutically acceptable carrier; (C) a pharmaceutical product comprising the inhalable form of the crystalline form of the invention in an inhalable form; And (D) an inhalation device comprising the crystalline form of the invention in an inhalable form.

Of course, the dosage of the agent of the invention used in the practice of the invention will depend, for example, on the particular condition to be treated, the desired effect, and the mode of administration. In general, a suitable daily dose for administration by inhalation is from about 0.0001 mg/kg to 30 mg/kg per patient, usually from 0.01 mg/kg to 10 mg/kg, and a suitable daily dose for oral administration is about 0.01 mg/kg. Up to 100 mg/kg.

The present invention further provides anhydrous pharmaceutical compositions and dosage forms comprising the crystalline form of the invention as an active ingredient, as water may promote degradation of certain compounds.

The anhydrous pharmaceutical compositions and dosage forms of the present invention can be prepared using anhydrous or low moisture containing ingredients and low moisture or low moisture conditions. Anhydrous pharmaceutical compositions can be prepared and stored to maintain their anhydrous character. Thus, anhydrous compositions are packaged using known materials to prevent exposure to water so that they can be included in a suitable formulation set. Examples of suitable packaging include, but are not limited to, hermetic sealing foils, plastics, unit dose containers (eg, vials), blister packs, and strip packs.

The invention further provides pharmaceutical compositions and dosage forms comprising one or more agents which reduce the rate of decomposition of the crystalline form of the invention as an active ingredient. Such agents referred to herein as "stabilizers" include, but are not limited to, antioxidants (e.g., ascorbic acid), pH buffers or salt buffers, and the like.

The crystalline form of the invention can be administered simultaneously with, or before or after administration of one or more additional therapeutic agents. The crystalline forms of the invention may be administered separately by the same or different routes of administration, or with other agents in the same pharmaceutical composition.

In one embodiment, the invention provides a product comprising a crystalline form of the invention and at least one other therapeutic agent in a combined formulation for simultaneous, separate or sequential use in therapy. In one embodiment, the therapy is a treatment for a disease or disease mediated by blocking epithelial sodium channels condition. The product provided in the combination preparation comprises a crystalline form of the invention comprising the crystalline form of the invention and the therapeutic agent(s) in the same pharmaceutical composition, or in a separate form (for example in the form of a kit) and another therapeutic treatment(s) Composition of the agent.

In one embodiment, the invention provides a pharmaceutical composition comprising a crystalline form of the invention and another therapeutic agent(s). Optionally, the pharmaceutical composition may comprise a pharmaceutically acceptable excipient as described above.

In one embodiment, the invention provides a kit comprising two or more separate pharmaceutical compositions, wherein at least one pharmaceutical composition comprises a crystalline form of the invention. In an embodiment, the kit includes means for separately retaining the compositions, such as a container, a discrete bottle, or a discrete foil package. Examples of such kits are blister packs, which are commonly used for packaged lozenges, capsules, and the like.

The kits of the invention can be used to administer different dosage forms (e.g., oral and parenteral dosage forms), to administer separate compositions at different dosage intervals, or to incrementally increase the dosage of the individual compositions relative to each other. To aid compliance, the kits of the present invention typically include instructions for administration.

In particular, the crystalline forms of the invention exhibit advantageous stability characteristics in human plasma. In providing compounds which exhibit advantageous stability characteristics in human plasma, the present invention also provides compounds which have improved pharmacokinetics to effectively block epithelial sodium channels (ENaC).

In the combination therapies of the invention, the crystalline forms of the invention and other therapeutic agents may be produced and/or formulated by the same or different manufacturers. Furthermore, the crystalline form of the invention and other therapeutic agents can be: (i) prior to the delivery of the combination product to the physician (eg, when present in a kit comprising the crystalline form of the invention and another therapeutic agent); Pre-administered by the physician itself (or under the direction of a physician); (iii) brought into the combination therapy by the patient itself, for example, during sequential administration of the compound of the invention and another therapeutic agent.

Accordingly, the present invention provides the use of the crystalline form of the present invention for use in the treatment of A disease or condition mediated by an epithelial sodium channel, wherein the agent is prepared for administration with another therapeutic agent. The invention also provides the use of another therapeutic agent for treating a disease or condition mediated by blocking epithelial sodium channels, wherein the agent is administered with a crystalline form of the invention.

The invention also provides a crystalline form of the invention for use in a method of treating a disease or condition mediated by epithelial sodium channels, wherein the crystalline form of the invention is prepared for administration with another therapeutic agent. The invention also provides another therapeutic agent for treating a disease or condition mediated by blocking epithelial sodium channels, wherein the additional therapeutic agent is prepared for administration with a crystalline form of the invention.

The invention also provides the use of a crystalline form of the invention for treating a disease or condition mediated by blocking epithelial sodium channels, wherein the patient has been previously treated (e.g., within 24 hours) with another therapeutic agent. The invention also provides the use of another therapeutic agent for treating a disease or condition mediated by blocking epithelial sodium channels, wherein the patient has previously received a crystalline form of the invention (e.g., within 24 hours).

In one embodiment, the additional therapeutic agent is selected from the group consisting of anti-inflammatory, bronchodilatory, antihistamine, decongestive, and antitussive drugs, particularly in cystic fibrosis or in obstructive or inflammatory respiratory diseases (eg, In the treatment of those mentioned, for example, as therapeutically potentiating agents of such drugs or as a means of reducing the required dose or potential side effects of such drugs.

Accordingly, the present invention includes the combination of the epithelial sodium channel blocker of the present invention and the following materials as another embodiment: a penetrating agent (high saline, dextran, mannitol, xylitol), a CFTR functional modifier ( Both the wild type and the mutant type (correcting agent and synergist), for example, are as described in the following: WO2007/021982, WO2006/099256, WO2006/127588, WO2004/080972, WO2005/026137, WO2005/035514 WO2005/075435, WO2004/111014, WO2006/101740, WO2004/110352, WO2005/120497 and US2005/0176761, anti-inflammatory, bronchiectasis, antihistamine, antitussive, antibiotic or DNase drug, the epithelial sodium channel blocker and the drug system are present in the same or different pharmaceutical compositions.

Suitable modifiers for the CFTR function include CFTR synergists, specifically the compound VX-770 of the formula below.

Suitable antibiotics include macrolide antibiotics, giant, e.g., tobramycin (tobramycin) (TOBI ^TM).

Suitable drugs include α deoxyribose enzyme (Pulmozyme ^TM) (recombinant human deoxyribonuclease of high purity solution I (rh) of) selectively dissociated DNA. Alpha deoxyribosylase is used to treat cystic fibrosis.

Other useful combinations of epithelial sodium channel blockers and anti-inflammatory drugs contain chemokine receptors (eg, CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-). 7. Antagonists of CCR-8, CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5), especially CCR-5 antagonists, such as Schering-Plough antagonists SC-351125, SCH-55700 and SCH- D; Takeda antagonist, such as N -[[4-[[[6,7-dihydro-2-(4-methylphenyl)-5H-benzo-cyclohepten-8-yl]carbonyl]amine Phenyl]-phenyl]-methyl]tetrahydro- N,N -dimethyl- 2H -pyran-4-ammonium chloride (TAK-770); and described in USP 6,166,037 (particularly the scope of patent application 18 And 19th), WO00/66558 (particularly the scope of application of claim 8), WO 00/66559 (particularly the scope of application of claim 9), WO 04/018425 and WO 04/026873 CCR-5 antagonists.

Suitable anti-inflammatory drugs include steroids, in particular, glucocorticosteroids such as budesonide, beclomethasone dipropionate, fluticasone propionate, ciclesonide or citric acid. Mometasone furoate, or the steroids described below: WO 02/88167, WO 02/12266, WO 02/100879, WO 02/00679 (particularly examples 3, 11, 14, 17, 19, 26, 34, 37, 39, 51, 60, 67, 72, 73, 90, 99 and 101), WO 03/35668, WO 03/48181, WO 03/62259, WO 03/64445, WO 03/72592, WO 04/39827 and WO 04/66920; non-steroidal glucocorticoid receptor agonists, for example, as described in the following: DE 10261874, WO 00/00531, WO 02/10143, WO 03/82280, WO 03/ 82787, WO 03/86294, WO 03/104195, WO 03/101932, WO 04/05229, WO 04/18429, WO 04/19935 and WO 04/26248; LTD4 antagonists, such as montelukast and Zafirlukast; PDE4 inhibitors such as cilomilast (Ariflo ^® GlaxoSmithKline), Roflumilast (Byk Gulden) ), V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering-Plough), Arofylline (Almirall Prodesfarma), PD189659/PD168787 (Parke-Davis), AWD-12- 281 (Asta Medica), CDC-801 (Celgene), SelCID (TM) CC-10004 (Celgene), VM554/UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo) and the following The disclosures of which are: WO 92/19594, WO 93/19749, WO 93/19750, WO 93/19751, WO 98/18796, WO 99/16766, WO 01/13953, WO 03/104204, WO 03/104205, WO 03/39544, WO 04/000814, WO 04/000839, WO 04/005258, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/018431, WO 04/018449, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/019944, WO 04/019945, WO 04/045607, and WO 04/037805; adenosine A2B receptor antagonists, for example, Equivalent to those described in WO 02/42298; and β-2 adrenergic receptor agonists, such as albuterol (salbutamol), metaproterenol, terbutaline , salmeterol, fenoterol, Procatorol, and in particular formoterol, carmoterol and pharmaceutically acceptable salts thereof, and compounds of formula (I) of WO0075114 (in free or salt or solvate form) The document is incorporated herein by reference, the preferred compounds of the examples, especially the compounds of the formula: It corresponds to indacaterol and its pharmaceutically acceptable salts, and the compound of formula I of WO 04/16601 (in the form of a free or salt or solvate), and the compounds of the following: EP 1440966, JP 05025045, WO 93/18007, WO 99/64035, USP 2002/0055651, WO 01/42193, WO 01/83462, WO 02/66422, WO 02/70490, WO 02/76933, WO 03/24439, WO 03/ 42160, WO 03/42164, WO 03/72539, WO 03/91204, WO 03/99764, WO 04/16578, WO 04/22547, WO 04/32921, WO 04/33412, WO 04/37768, WO 04/ 37773, WO 04/37807, WO 04/39762, WO 04/39766, WO 04/45618, WO 04/46083, WO 04/80964, WO 04/108765 and WO 04/108676.

Suitable bronchodilators include anticholinergic or antimuscarinic agents, in particular, ipratropium bromide, oxitropium bromide, tiotropium salts and CHF 4226 (Chiesi), and glycopyrrolate, and the same as those described in the following: EP 424021, USP 3,714,357, USP 5,171,744, WO 01/04118, WO 02/00652, WO 02/51841, WO 02/53564, WO 03/00840, WO 03/33495, WO 03/53966, WO 03/87094, WO 04/018422 and WO 04/05285.

Suitable dual anti-inflammatory and bronchodilators include dual beta-2 adrenergic receptor agonists/muscarinic antagonists, for example, as disclosed in USP 2004/0167167, WO 04/74246, and WO 04/74812.

Suitable antihistamines include cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, loratadine ( Loratidine), desloratidine, diphenhydramine, and fexofenadine hydrochloride, activastine, astemizole, azelastine Azelastine), ebastine, epinastine, mizolastine, and tefenadine, and the same as JP 2004107299, WO 03/099807, and WO 04/026841 Revealed.

The term "pharmaceutically acceptable carrier" as used in the present invention includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (eg, antibacterial agents, known to those skilled in the art). Antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, pharmaceuticals, pharmaceutical stabilizers, binders, excipients, disintegrating agents, lubricants, sweeteners, flavoring agents, dyes, and the like and Combinations thereof (for example, see Remington's Pharmaceutical Sciences, 18th ed., Mack Printing, 1990, pages 1289 to 1329). In addition to any conventional carrier that is incompatible with the active ingredient, the invention also encompasses its use in therapeutic or pharmaceutical compositions.

The term "therapeutically effective amount" of the crystalline form of the invention means that the individual is allowed to produce a biological or medical response (eg, to reduce or inhibit enzyme or protein activity), or to ameliorate symptoms, alleviate the condition, slow or delay disease progression, or prevent disease, etc. The amount of the crystalline form of the invention. In one non-limiting embodiment, the term "therapeutically effective amount" refers to the following when administered to an individual Effective amounts of the crystalline form of the invention: (1) at least partially alleviate, inhibit, prevent and/or ameliorate (i) mediated by epithelial sodium channels or (ii) associated with epithelial sodium channel activity, or (iii) epithelial sodium Channel activity (normal or abnormal) is a condition or disorder or disease characterized; or (2) reducing or inhibiting epithelial sodium channel activity. In another non-limiting embodiment, the term "therapeutically effective amount" refers to the invention that is effective to at least partially reduce or inhibit the activity of the epithelial sodium channel when administered to a cell, or tissue or non-cellular biological material, or a medium. The amount of crystalline form.

The term "individual" as used herein refers to an animal. Typically, the animal is a mammal. For example, a subject also refers to primates (eg, humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, and the like. In some embodiments, the system is primate. In still other embodiments, the system is human.

The term "inhibiting, inhibiting, or inhibiting" as used herein refers to alleviating or suppressing a given condition, symptom, or condition, or disease, or significantly reducing the baseline activity of a biological activity or process.

In one embodiment, the term "treat, treating, or treating" as used herein refers to amelioration of the disease or condition (ie, slowing or preventing or reducing the progression of the disease or at least one of its clinical symptoms). . In another embodiment, "treat, treating, or treating" refers to mitigating or improving at least one physical parameter that includes physical parameters that such patients may not be able to perceive. In still another embodiment, "treat, treating, or treating" refers to physically modulating a disease or condition (eg, stabilizing a sensible symptom) or physiologically modulating a disease or condition (eg, stabilizing physics) Parameter) or both. In yet another embodiment, "treat, treating, or treating" refers to preventing or delaying the onset or progression or progression of a disease or condition.

If the individual used herein would benefit from treatment in terms of biology, medicine, or quality of life, the individual "needs" the treatment.

The term "a", as used herein, is used in the context of the present invention (especially in the context of the claims), unless otherwise indicated herein or clearly contradicted by the context. "The" and similar terms are understood to cover both the singular and the plural.

experiment General conditions:

Mass spectrometry can be run on the LCMS system using electrospray free.

The LCMS systems are an Agilent 1100 HPLC/Micromass Platform mass spectrometer combination, or an Agilent 1200 HPLC/Agilent 6130 Quadropole mass spectrometer combination, or a Waters Acquity UPLC with an SQD mass spectrometer. [M+H] ⁺ means the molecular weight of a single isotope.

NMR spectra were run on an open access Bruker AVANCE 400 NMR spectrometer using ICON-NMR. The spectra were measured at 298 K and referenced using solvent peaks. Due to the extremely wide nature of its exchangeable resonance, some protons were not directly observed.

Analytical HPLC conditions are as follows, unless otherwise indicated:

abbreviation:

Preparation 1: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl ester

The title compound can be prepared by Method A or B.

Method A:

To 3-(2-dipropylaminecarbomethoxycarbonyl-ethylaminesulfonyl)-benzoic acid (prepared as in Preparation 2; 6.1 g, 12.60 mmol) in THF (50 mL) Water (25 ml), N-methylmorpholine (7 ml, 63 mmol) and HOBt hydrate (2.9 g, 18.9 mmol) were sequentially added to the stirred solution. Maintain internal temperature at At 20 ° C. Addition of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid [1,3,8-triaza-spiro[4.5]indole-(2E)-ylidene]-nonylamine hydrochloride ( WO09/074575, Example 38, page 123) (purity 65%, 6.3 g, 12.6 mmol) and stirred until a clear solution was formed. EDCI.HCl (3.6 g, 18.9 mmol) was added and the mixture was stirred at room temperature for 24 h. 2-MeTHF (200 ml) and a 2% aqueous Na ₂ CO ₃ solution (150 ml) were added to the reaction mixture. The layers were separated and the aqueous phase was washed with additional 2-MeTHF (100 mL). The organic layers were washed with _2% Na 2 CO ₃ solution (200ml) and water (2 × 200ml). Acetonitrile (100 ml) was added and the solution was concentrated to a volume of 70 ml at 30 °C. Acetonitrile (300 ml) was added and the solution was again concentrated to a volume of 150 ml at 30 °C. The solution was heated to 50 ° C and maleic acid (1.62 g) was added to the resulting solution. An off-white precipitate formed immediately and the temperature was cooled to room temperature over 1 h. The solid was collected by filtration to give 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino) as a maleate salt. -1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester; DCM (200 ml) and A 2% aqueous Na ₂ CO ₃ solution (200 ml) was stirred until the solid was completely dissolved. The organic layer was separated, washed with w~~~ LC-MS 722.1 [M + H ] +, method ^{(i); 1 H NMR (} 400MHz, DMSO-d6) δ 9.12-7.57 (4H, br), 7.88 (1H, m), 7.77 (1H, m), 7.70 (1H, m), 7.68 (1H, m), 7.05-6.50 (2H, br s), 6.95-6.20 (1H, br s), 4.73 (2H, s), 3.81-3.39 (2H, m), 3.61-3.31 (2H, m), 3.43 (2H, br s), 3.15-3.11 (4H, m), 3.04 (2H, t), 2.51 (2H, t), 1.79-1.69 (m, 4H), 1.51 -1.43 (4H, m), 0.84 (3H, t), 0.78 (3H, t)

Method B:

Stirring solution to 3-(2-dipropylamine-methylhydrazolylmethoxycarbonyl-ethylaminesulfonyl)-benzoic acid (Int.AA) (6.1 g, 12.60 mmol) in THF (50 mL) Water (25 ml), N-methylmorpholine (7 ml, 63 mmol) and HOBt hydrate (2.9 g, 18.9 mmol) were sequentially added. Maintain internal temperature at At 20 ° C. Addition of 3,5-diamino-6-chloro-pyrazine-2-carboxylic acid [1,3,8-triaza-spiro[4.5]indole-(2E)-ylidene]-nonylamine hydrochloride ( WO09074575, Example 38, page 123) (purity 65%, 6.3 g, 12.6 mmol) and stirred until a clear solution formed. EDCI.HCl (3.6 g, 18.9 mmol) was added and the mixture was stirred at room temperature for 24 h. 2-MeTHF (200 ml) and a 2% aqueous Na ₂ CO ₃ solution (150 ml) were added to the reaction mixture. The layers were separated and the aqueous phase was washed with additional 2-MeTHF (100 mL). The organic layers were washed with _2% Na 2 CO ₃ solution (200ml) and water (2 × 200ml). Acetonitrile (100 ml) was added and the solution was concentrated to a volume of 70 ml at 30 °C. Acetonitrile (300 ml) was added and the solution was again concentrated to a volume of 150 ml at 30 °C. The solution was heated to 50 ° C and maleic acid (1.62 g) was added to the resulting solution. An off-white precipitate formed immediately and the temperature was cooled to room temperature over 1 h. The solid was collected by filtration to give 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino) as a maleate salt. -1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester; DCM (200 ml) and A 2% aqueous Na ₂ CO ₃ solution (200 ml) was stirred until the solid was completely dissolved. The organic layer was separated, washed with w~~~ LC-MS 722.1 [M + H ] +, method ^{(i) .; 1 H NMR (} 400MHz, DMSO-d6) δ 9.12-7.57 (4H, br), 7.88 (1H, m), 7.77 (1H, m) , 7.70 (1H, m), 7.68 (1H, m), 7.05-6.50 (2H, br s), 6.95-6.20 (1H, br s), 4.73 (2H, s), 3.81-3.39 (2H, m) , 3.61-3.31 (2H, m), 3.43 (2H, br s), 3.15-3.11 (4H, m), 3.04 (2H, t), 2.51 (2H, t), 1.79-1.69 (m, 4H), 1.51-1.43 (4H, m), 0.84 (3H, t), 0.78 (3H, t)

Preparation 2: 3-(2-Dipropylamine-carbenylmethoxycarbonyl-ethylaminesulfonyl)-benzoic acid

The title compound can be prepared by Method A or B:

Method A:

Step 1 : 3-Benzyloxycarbonylamino-propionic acid dipropylamine-methylmethyl ester

Potassium carbonate (19.3 g, 139.9 mmol) was added to a solution of benzyloxycarbonylamino-propionic acid (22.3 g, 99.9 mmol) in DMF (150 mL). 2-Chloro-N,N-dipropyl-acetamide (17.7 g, 99.9 mmol) was added over 30 min and the reaction mixture was heated to 60 ° C and stirred for 2.5 h. The reaction mixture was cooled to room temperature and diluted with H.sub.2 (500 mL). The combined organic phases were washed with water (3.times..times.ssssssssssssssssssssssssssssssssssssssssssssssssssss LC-MS; 365.2 [M+H] ⁺ , (m).

Step 2 : 3-Amino-propionic acid dipropylamine-methylmethylmethyl trifluoroacetate

A solution of 3-benzyloxycarbonylamino-propionic acid dipropylamine methyl decyl methyl ester in isopropyl acetate (33.2 g, 91.0 mmol in total mass) was treated with TFA (7.05 ml, 92.0 mmol). 129.9 g of the solution) while maintaining the internal temperature at 20 ° C, followed by treatment with 10% Pd/C (3.3 g, humidity 50%) and stirring for 3.5 h under H ₂ atmosphere (3 atm) to give the 3-amine Base-dipropylamine-methylmethylmethyl trifluoroacetate. This solution was used in the next reaction without isolation.

Step 3 : 3-(2-Dipropylamine-mercaptomethoxycarbonyl-ethylaminesulfonyl)-benzyl benzoate

A solution of 3-amino-propionic acid dipropylamine methyl decyl methyl ester trifluoroacetate (25.4 g 73.8 mmol) in isopropyl acetate was cooled to 0 ° C with N-methylmorpholine (26.3 g , 221.5 mmol), water (40 ml) and DMAP (90.4 mg, 0.74 mmol). 3-Chlorosulfonyl-benzoic acid benzyl ester (24.1 g, 77.5 mmol) in isopropyl acetate (44 ml) was added and the mixture was stirred at 0 ° C to 5 ° C for 2 h. The layers were separated and diluted with saturated aqueous NaHCO ₃ (3 × 26ml) The organic phase was washed with water (10ml), washed with 1N HCl solution was adjusted to pH 6, and washed with brine. The resulting solution is concentrated to provide a solution of 3-(2-dipropylamine-methanecarbonylcarbonyl-ethylaminesulfonyl)-benzoic acid benzyl ester in isopropyl acetate which is not isolated. Used in other reactions. LC-MS; [M+H] ⁺ 505.1.

Step 4 : Synthesis of 3-(2-dipropylamine-mercaptomethoxycarbonyl-ethylaminesulfonyl)-benzoic acid

Treatment of 3-(2-dipropylamine-mercaptomethoxycarbonyl-ethylaminesulfonyl)-benzyl benzoate (5 g, 33.25 mmol) with 10% Pd/C (0.84 g, 50%) ) in isopropyl acetate () and the solution was 82ml, and stirred under H ₂ (3atm) overnight. The catalyst is removed by filtration. 10% Pd/C (1.68 g, 50% humidity) was added to the filtrate, and the reaction was stirred at H ₂ (3 atm) for 18 h. The catalyst was removed by filtration and 10% Pd/C (1.68 g, 50% humidity) was then added and the mixture was stirred at H ₂ (1 atm) for 18 h. The catalyst was removed by filtration and washed with isopropyl acetate (20 mL). The combined filtrate was concentrated in vacuo and heptane was added to the mixture and stirred at room temperature for 2 hr then stirred at -2 ° C for 4 h. The solid formed was collected by filtration and dried <RTI ID=0.0></RTI> to ^{<RTI ID} =0.0>

Method B:

Step 1: 3-Tertoxycarbonylamino-propionic acid dipropylamine methyl decyl methyl ester

Potassium carbonate (40.0g, 289mmol) to a stirred suspension of the Boc-Beta-Ala-OH ( 40.0g, 211mmol) in DMF (200ml) under N ₂ and was at 60 ℃. To this mixture was added 2-chloro-N,N-dipropyl-acetamide (36.7 g, 207 mmol) in DMF (75 mL). The reaction mixture was stirred at 60 ° C overnight. The reaction was cooled to room temperature and diluted with DCM (400 mL). The organic layer was separated and washed with brine (200ml), concentrated in vacuo and dried over MgSO _4, to give a pale yellow oil. To this oil was added n-heptane (500ml) (DMF of azeotrope), and concentrated in vacuo to give the title compound; LC-MS Rt 1.14min; 331.3 [M + H] +, Method 2minLC_vO03.

Step 2: 3-Amino-propionic acid dipropylamine-methylmethyl methyl ester

Under N ₂ to 3-tert-butoxycarbonyl-amino - propionic acid methyl dipropylamine acyl methyl ester (Step 1) (36.5g, 110mmol) in dry dioxane was cooled to stir the 4N HCl (18.12 ml, 597 mmol) in dioxane was added dropwise to the solution. The resulting mixture was allowed to warm to room temperature and stirred overnight. The solvent was removed in vacuo and the crude was taken in EtOAc (EtOAc) The resulting white precipitate was isolated by filtration and dried at 40 &lt Recrystallization from EtOAc, to give the title compound; LC-MS Rt 0.77min; 231.2 [M + H] +, Method 2minLC_v003.

Step 3: 3-(2-Dipropylamine-carbenylmethoxycarbonyl-ethylaminesulfonyl)-benzoic acid

(240ml) in the stirred solution was added DMAP (0.46 dipropylamine acid methyl acyl methyl ester (Step 2) (20.1g, 75mmol) in DCM - at 0 ℃ under N ₂ and the 3-amino g, 3.76 mmol) followed by TEA (38.8 mL, 278 mmol). The reaction mixture was treated with a solution of 3-chlorosulfonylbenzoic acid (16.6 g, 75 mmol) in DCM The mixture was stirred at 0 °C for 1 h and then warmed to room temperature for 1 h. Water (200 ml) was added and the pH was adjusted with 1N HCl (100 mL). The organic layer was separated, dried over MgSO ₄ By C18 reverse phase chromatography eluting embodiment purified using MeCN / water (1% HCl), to give the title product; LC-MS Rt 1.01min; 415.2 [M + H] +, Method 2minLC_v003.

Example 1: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form A

2.5 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-three at 50 °C Aza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl ester (prepared as prepared in Preparation 1; 3.47 mmol) is dissolved in 25 mL of acetonitrile And 1.5 mL of water. 409 mg of succinic acid (3.47 mmol) was added. The acid dissolves immediately and passes for 30 minutes. The clear solution was cooled to room temperature. Crystallization occurs at about 30 °C. The slurry was then stirred at room temperature for about 16 h. The crystals were collected by filtration. The filter cake was washed portionwise with 9 ml acetonitrile / water 95:5 v / v and dried at 50 ° C and about 10 mbar for 16 h.

Example 2: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form A

A mixture comprising succinic acid (0.50 g, 4.23 mmol) and acetone (20 g) was heated to 45 °C until a clear solution was formed and then filtered (0.2 [mu]m PTFE filter).

In a further reaction vessel, 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8- Triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethyl ester (Preparation 1, Method B) (3.00 g, 4.16 mmol) Acetone (30 g) was heated to 45 ° C until a clear solution was formed and then filtered (0.2 μm PTFE filter).

A solution of succinic acid (0.50 g, 4.23 mmol) in acetone (20 ml) was heated at 45 ° C for 1 h and then with 3-(3-{2-[(E)-3,5-diamino-6 -Chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine A portion of the solution of formazan methyl ester in acetone (1.62 g solution) was treated. 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[4.5 ] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethyl succinate seed crystal (prepared as prepared using the preparation of Example 1, 20 mg) in acetone (300 mg) The resulting mixture was treated with a suspension in the mixture and stirred at 45 ° C for 30 min. The remaining 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza is added over 5 h - a solution of spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylhydrazine methyl ester in acetone (31.38 g) was added to the mixture at 45 ° C Continue heating for 1 h. The suspension was cooled to 25 ° C over 1 h and stirred for an additional 1 h. The suspension was filtered on a glass frit and the filter cake was washed with acetone (2 x 5 g). The filter cake was dried at 50 deg.] C, to give the title compound; HPLC Rt 4.02min, method ^{ii; 1 H NMR (400MHz,} DMSO-d6) δ 7.87 (1H, m), 7.78 (1H, m), 7.69 (1H, m ), 7.68 (1H, m), 6.85 (2H, br s), 4.73 (2H, s), 3.84-3.20 (6H and water, wide hump), 3.17-3.09 (4H, m), 3.04 (2H, t ), 2.53 (2H in DMSO), 2.39 (4H, s), 1.80 (2H, br s), 1.70 (2H, br s), 1.55-1.37 (4H, m), 0.85 (3H, t), 0.78 (3H, t) (Note: due to the extremely wide nature of some exchangeable resonances, the two exchangeable succinate protons and 3 to 4 acidic NH resonances are not directly observed; melting temperature T _m (DSC) = 149 ° C.

Example 3: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form A

3 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[ 4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl succinate (prepared as shown in Example 1 or 2) is loaded to be equipped with an anchor Agitator (150 rpm), condenser, dosing unit and turbidity probe in a 100 ml reactor. 20 g of acetone/water mixture (92.5/7.5 m/m) was added and the whole was heated to 50 ° C and stirred until all dissolved. The reaction mixture was then cooled to 45 °C. 30 mg seed crystals (prepared as shown in Example 1 or 2) suspended in 300 mg of acetone were added. The reaction mixture was then cooled to 25 ° C over 1 hour. The whole was stirred for 3.5 hours. Then 28 ml of EtOAc was added over 2 hours. The reaction mixture was further cooled to 0 ° C over 1 hour and then the whole was stirred overnight. The reaction mixture was filtered through a glass filter and then the obtained solid was washed twice with 5 ml of acetone and dried at 40 ° C and 1 mbar for 24 hours.

Example 4: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form B

3 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-) in a 100 mL glass reactor equipped with an anchor stirrer (150 rpm) at 50 °C 2-carbonylimino]-1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester (can be as Prepared in Preparation 1) dissolved in 22.7 g of acetone. 5 mL of a succinic acid solution (2.94% by weight in acetone) was added to the reactor. 10 mg of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[ 4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl ester succinate crystalline form A (prepared as in Example 1 or 2) suspended in 0.2 g In acetone, ultrasonic treatment was carried out and added to the reactor. Another 40 mL of succinic acid solution (2.94% by weight in acetone) was slowly added to the reactor over 20 h. Thereafter, the mixture was cooled to 20 ° C and then further stirred for 19 h. The resulting suspension was filtered and the filter cake was washed twice with 10 g of acetone at room temperature.

Example 5: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form B

3 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-) in a 100 mL glass reactor equipped with an anchor stirrer (150 rpm) at 50 °C 2-carbonylimino]-1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester (Prepared as prepared in Preparation 1) and 0.5 g of succinic acid were dissolved in a mixture of 28.5 g of acetone and 1.5 g of water. Thereafter, the solution was cooled to 20 °C. 7 mg of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[ 4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl succinate crystal form A (prepared as in Example 1 or 2) suspended in 0.18 g In acetone/water 95:5 (v/v), ultrasonic treatment was carried out and added to the reactor. The mixture was stirred at 20 ° C for 15 hours and then cooled to 0 ° C in 10 min. The resulting suspension was filtered and the filter cake was washed with 10 g of acetone/water 95/5 (v/v) at room temperature.

Example 6: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form B

0.536 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino] in a magnetically stirred glass vial at 50 °C -1,3,8-triaza-spiro[4.5]decane-8-carbonyl}- Phenylsulfonylamino)-propionic acid dipropylamine methyl decyl methyl ester (prepared as prepared in Preparation 1) and 0.12 g of succinic acid were dissolved in 15.25 g of 2-propanol. The solution was cooled to 20 ° C and then stirred at this temperature overnight. The resulting suspension was filtered.

Example 7: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form B

0.535 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino] in a magnetically stirred glass vial at 60 °C -1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethyl ester (can be prepared as in Preparation 1) And 0.12 g of succinic acid was dissolved in 7.27 g of methyl ethyl ketone. The solution was cooled to 20 ° C and then stirred at this temperature overnight. The resulting suspension was filtered.

Example 8: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form C

3 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-) in a 100 mL glass reactor equipped with an anchor stirrer (150 rpm) at 60 °C 2-carbonylimino]-1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl ester (Can be prepared as in Preparation 1) dissolved in 80.3 g of ethanol. 25 mL of a succinic acid solution (2.94% by weight in ethanol) was added to the solution, followed by the addition of 2.5 g of water. The resulting mixture was cooled to 30 °C. 5 mg of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[ 4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl ester succinate seed crystal (Form A) (can be prepared by the preparation of Example 1 or 2) ) suspended in 0.1 g of ethanol, subjected to ultrasonic treatment, and added to the reactor. The resulting mixture was stirred for 96 h (150 rpm). Thereafter, the crystals were collected by filtration. The filter cake was dried at 40 ° C and 30 mbar for 24 hours.

Example 9: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triazole Hetero-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: preparation of crystalline form C

1 g of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3 in a glass vial at 50 °C, 8-Triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine methyl decyl methyl ester (prepared as prepared in Preparation 1) and 0.18 g amber The acid was dissolved in 7.97 g of methanol. The solution was cooled to 0 ° C and then stirred at this temperature for 24 hours. Thereafter, the crystals were collected by filtration and dried at 60 ° C and 1 mbar for 3 hours. Obtained a pale white to pale yellow crystal form C.

Example 10: 3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro [4.5] decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate: Preparation of crystalline form C 10.1 Preparation of crystals:

In a 100 ml reactor equipped with an anchor stirrer (150 rpm), condenser, dosing unit and turbidity probe, 6 g of 3-(3-{2-[(E)-3,5-diamino- 6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropyl Aminomethylmercaptomethyl succinate (prepared as shown in Example 1 or 2) was suspended in 27 g of an acetone/water mixture (92.5/7.5 m/m). The suspension was heated to 50 ° C until the material dissolved. Add 40 mg of 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino] in 600 mg of acetone after ultrasonic treatment for 5 minutes] -1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate (as in Example 9) Prepared as shown) and stirred as a whole for 6 hours. 76 ml EtOAc was added over 4 hours and then the reaction mixture was cooled to 25 °C over 2 hr. The precipitate was filtered and washed twice with 10 ml of acetone and then dried overnight at 60 ° C and 10 mbar. The resulting solid was then micronized using a spiral jet mill at 5 bar.

10.2 Preparation of crystalline form C:

3g 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-three Aza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate (prepared as shown in Example 1 or 2) Charge into a 100 ml reactor equipped with an anchor stirrer (150 rpm), condenser, dosing unit and turbidity probe. 29.5 g of acetone/water mixture (92.5/7.5 m/m) was added and the whole was heated to 50 ° C and stirred until all dissolved. 90 mg seed crystals (see 10.1) suspended in 300 mg of acetone were added and stirred for 4 hours. 38 ml of EtOAc was added over 4 hours and then the reaction mixture was stirred for 2 h then cooled to 25 °C over 2 h. The whole was stirred overnight and then filtered through a glass filter. The solid obtained was then washed twice with 5 ml of acetone and dried at 40 ° C and 10 mbar for 24 hours.

3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza-spiro[4.5] Characterization of the crystalline form of decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate:

All X-ray powder diffraction patterns were recorded in a reflective mode.

Type A:

a) X-ray powder diffraction

a1) using CuKα radiation (λ = 1.5418Å) x- ray powder diffraction pattern recorded on a Bruker ^TM D8 diffractometer. The X-ray diffraction pattern of Example 3 thus determined is shown in Figure 1 and is shown in Table 1 below as the most significant line of reflection. The error limit of the 2-θ angle is ±0.2°.

a2) using CuKα radiation (λ = 1.5418Å) x- ray powder diffraction pattern recorded on a Bruker ^TM D8 diffractometer. The X-ray diffraction pattern of Example 1 or 2 thus determined is shown in Figure 4 and is shown in Table 2 below as the most significant line of reflection. The error limit of the 2-θ angle is ±0.2°.

b) Elemental analysis: water content (Karl Fischer titration): <0.2% m/m

The experimental data is consistent with 3-(3-{2-[(E)-3,5-diamino-6-chloro-pyrazine-2-carbonylimine) 1,1,3,8-triaza-spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethyl ester and succinic acid Expectation of 1:1 salt.

c) Differential Scanning Calorimetry (DSC): DSC traces were recorded on a Perkin Elmer Diamond DSC instrument (Perkin Elmer, model BO14-3018) with an aluminum pan; heating rate 20 K/min, temperature range: 30 ° C to 250 ° C .

Figure 2 shows the DSC trace of version A. Melting _{endotherm: T ℃, △} H = 76.8J / g.

Figure 5 shows the DSC trace of version A. Melting _{endotherm: T ℃, △} H = 74.5J / g.

c) Infrared Spectroscopy: Figure 3 shows the infrared spectrum of Form A (Example 3) and recorded on a Bruker Alpha instrument in an attenuated total reflectance (ATR) mode. The following parameters were measured: range 400 cm ^-1 to 4000 cm ^-1 , resolution 2 cm ^-1 , number of scans 64, speed 7.5 kHz, apodization method: Blackman-Harris 3-term. The identified main peaks were 1621.5 cm ^-1 , 1157.5 cm ^-1 and 1080.7 cm ^-1 .

Type B:

X- ray powder diffraction: using CuKα radiation (λ = 1.5418Å) x- ray powder diffraction pattern recorded on a Bruker ^TM D8 diffractometer. The X-ray diffraction pattern thus determined is shown in Fig. 6 and is shown in Table 4 below as the reflection line of the most important line. The error limit of the 2-θ angle is ±0.2°.

Type C:

a) X-ray powder diffraction:

a1) using CuKα radiation (λ = 1.5418Å) x- ray powder diffraction pattern recorded on a Bruker ^TM D8 diffractometer. The X-ray diffraction pattern thus determined is shown in Fig. 7 and is shown in Table 5 below as the reflection line of the most important line. The error limit of the 2-θ angle is ±0.2°.

a2) using CuKα radiation (λ = 1.5418Å) x- ray powder diffraction pattern recorded on a Bruker ^TM D8 diffractometer. The X-ray diffraction pattern of Example 8 or 9 thus determined is shown in Figure 10 and is shown in Table 6 below as the most significant line of reflection. The error limit of the 2-θ angle is ±0.2°.

The laboratory temperature varied between 21 ° C and 26 ° C during the recording of XRPD data.

b) Differential Scanning Calorimetry (DSC): DSC traces were recorded on a Perkin Elmer Diamond DSC instrument (Perkin Elmer, model BO14-3018) with aluminum pan; heating rate 20 K/min, temperature range: 30 ° C to 250 ° C .

Figure 8 shows the DSC trace of Form C (Example 10). Melting _{endotherm: T ℃, △} H = 102.2J / g.

Figure 11 shows the DSC trace of version C (Examples 8, 9). Melting _{endotherm: T ℃, △} H = 91.8J / g.

c) Infrared Spectroscopy: Figure 9 shows the infrared spectrum of Form C (Example 10) and recorded on a Bruker Alpha instrument in an attenuated total reflectance (ATR) mode. The following parameters were measured: range 400 cm ^-1 to 4000 cm ^-1 , resolution 2 cm ^-1 , number of scans 64, speed 7.5 kHz, apodization method: Blackman-Harris 3-term. The peak identified based 1626.5cm ^-1, 1155cm ^-1 and 1084.7cm ^-1.

Claims (12)

3-(3-{2-[(E)-3,5-Diamino-6-chloro-pyrazine-2-carbonylimino]-1,3,8-triaza- a crystalline form of spiro[4.5]decane-8-carbonyl}-benzenesulfonylamino)-propionic acid dipropylamine-methylmethylmethyl succinate, It consists essentially of Form C, characterized in that the x-ray powder diffraction pattern has a 2-theta value comprising 8 or more selected from the group consisting of: 7.0° ± 0.2 at a temperature of 21 ° C to 26 ° C. 10.6°±0.2, 14.3°±0.2, 18.2°±0.2, 18.6°±0.2, 19.2°±0.2, 21.2°±0.2, 21.8°±0.2, 24.7°±0.2, 29.0°±0.2 and 31.5°±0.2 . A crystalline form as claimed in claim 1, wherein the version C comprises less than 10% by weight of any other physical form of the compound. The crystalline form of claim 1, wherein the x-ray powder diffraction spectrum is substantially the same as the x-ray powder diffraction spectrum shown in FIG. The crystalline version of claim 1 wherein the differential scanning calorimetry thermogram is substantially identical to the differential scanning calorimetry thermogram shown in FIG. A pharmaceutical composition comprising the crystalline form of any one of claims 1 to 4 and a pharmaceutically acceptable carrier or diluent. A pharmaceutical composition according to claim 5, which is combined with one or more other active ingredients. The pharmaceutical composition of claim 5 or 6, which is in an inhalable form. A crystalline form according to any one of claims 1, 3 and 4 for use in the treatment of a respiratory disease. A crystalline form according to any one of claims 1, 3 and 4 for use in the treatment of cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory infection And lung cancer. Use of the crystalline form of any one of claims 1 to 4 for the manufacture of a medicament for the treatment of a respiratory disease. Use of a crystalline form according to any one of claims 1 to 4 for the manufacture of cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD) , asthma, respiratory infections and lung cancer agents. An inhalation device comprising the crystalline form of any one of claims 1 to 4 and adapted to deliver the crystalline form by pulmonary administration.